Week of 7/5

From 2010.igem.org

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==July 5th==
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 +
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*The university was closed due to the holiday on July 4th.
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 +
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==July 6th==
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 +
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*Back to work!
 +
 +
*Tomorrow:
 +
  - Retransform:
 +
        o All lux promoters from kit plates.
 +
 
 +
*Grow for tomorrow from stock:
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        - R0061
 +
        - J24679
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        - R1062
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        - K112022
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        - K091107
 +
        - K091146
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        - K145150
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*Cleaned cuvettes for transformations.
 +
 +
*Worked on Human Practices survey.
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==July 7th==
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*Flasks minipreped:
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      - J24679
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      - R1062
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      - K112022
 +
      - K091107
 +
      - K091146
 +
      - K145150
 +
 +
*Sent to sequencing:
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  1) P0451
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  2) K112022
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  3) J24679
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  4) K091146
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  5) K145150
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  6) R0010
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  7) R1062
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  8) K091107
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 +
 +
*Digests:
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  - J24679 (ES+phosphatase)
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  - P0451  (XP+phosphatase)
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  - K274100(ES+phosphatase)
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  - S01414 (ES+phosphatase)
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  - K274100(XP+phosphatase)
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  - B1002  (EX)
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  - B1006  (EX)
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  - R0010  (SP)
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  - B1002  (EX)
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  - B1006  (EX)
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  - J23100 (SP)
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*Gel order:
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  - Ladder
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  - B1006
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  - J23100
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  - R0010
<center>[[Image:7-7-10.jpg]]</center>
<center>[[Image:7-7-10.jpg]]</center>
 +
 +
All three bands appeared around the base pairs that we wanted then at so the 3 bands were cut out and then gel purified.
 +
 +
*Ordered 3 primers.
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 +
*Ligations:
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  Insert          Vector
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1) J24679          B1006
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2) P0451          R0010
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3) K274100        B1006
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4) S01414          B1006
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5) K274100        J23100
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 +
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==July 8th==
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 +
*Colony PCR:
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  1) P0451+R0010 A-D
 +
  2) K274100+B1006 A-D
 +
  3) K274100+J23100 A-D
 +
  4) J24679+B1006 A-D
 +
 +
 +
*Digest:
 +
  - J24679 (ES+phosphatase)
 +
  - K274100(ES)
 +
  - R0010 (SP)
 +
  - B1002 (EX)
 +
  - K112022 (XP)
 +
  - K112808 (XP)
 +
*Gel order:
 +
  Upper-digests
 +
    - Ladder
 +
    - K274100
 +
    - K112808
 +
    - K1120222
 +
    - B1002
 +
  Lower-PCR
 +
    - Ladder
 +
    - 1 A-F
 +
    - 4 A-E
<center>[[Image:7-8-10.jpg]]</center>
<center>[[Image:7-8-10.jpg]]</center>
 +
*We forgot to put the Taq polymerase and so as you can see, nothing showed up in the gel.
<center>[[Image:7-9-10.jpg]]</center>
<center>[[Image:7-9-10.jpg]]</center>

Revision as of 18:41, 4 August 2010

Contents

July 5th

  • The university was closed due to the holiday on July 4th.


July 6th

  • Back to work!
  • Tomorrow: 
  - Retransform:
       o All lux promoters from kit plates.
  • Grow for tomorrow from stock: 
       - R0061
       - J24679
       - R1062
       - K112022
       - K091107
       - K091146
       - K145150
  • Cleaned cuvettes for transformations.
  • Worked on Human Practices survey.


July 7th

  • Flasks minipreped: 
      - J24679
      - R1062
      - K112022
      - K091107
      - K091146
      - K145150
  • Sent to sequencing: 
 1) P0451
 2) K112022
 3) J24679
 4) K091146
 5) K145150 
 6) R0010
 7) R1062
 8) K091107


  • Digests: 
  - J24679 (ES+phosphatase)
  - P0451  (XP+phosphatase)
  - K274100(ES+phosphatase)
  - S01414 (ES+phosphatase)
  - K274100(XP+phosphatase)
  - B1002  (EX)
  - B1006  (EX)
  - R0010  (SP)
  - B1002  (EX)
  - B1006  (EX)
  - J23100 (SP)


  • Gel order: 
  - Ladder
  - B1006
  - J23100
  - R0010
7-7-10.jpg

All three bands appeared around the base pairs that we wanted then at so the 3 bands were cut out and then gel purified.

  • Ordered 3 primers.
  • Ligations: 
  Insert          Vector

1) J24679 B1006 2) P0451 R0010 3) K274100 B1006 4) S01414 B1006 5) K274100 J23100


July 8th

  • Colony PCR: 
 1) P0451+R0010 A-D
 2) K274100+B1006 A-D 
 3) K274100+J23100 A-D
 4) J24679+B1006 A-D


  • Digest: 
 - J24679 (ES+phosphatase)
 - K274100(ES)
 - R0010 (SP)
 - B1002 (EX)
 - K112022 (XP)
 - K112808 (XP)


  • Gel order: 
 Upper-digests
    - Ladder
    - K274100
    - K112808
    - K1120222
    - B1002
 Lower-PCR
    - Ladder
    - 1 A-F
    - 4 A-E
7-8-10.jpg
  • We forgot to put the Taq polymerase and so as you can see, nothing showed up in the gel.


7-9-10.jpg
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